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Role of Saccharomyces cerevisiae TAN1 (tRNA acetyltransferase) in eukaryotic initiation factor 2B (eIF2B)-mediated translation control and stress response
| Content Provider | Semantic Scholar |
|---|---|
| Author | Sharma, Sonum Sourirajan, Anuradha Dev, K. N. Prabhu |
| Copyright Year | 2017 |
| Abstract | Eukaryotic initiation factor 2B (eIF2B) controls the first step of translation by catalyzing guanine nucleotide exchange on eukaryotic initiation factor 2 (eIF2). Mutations in the genes encoding eIF2B subunits inhibit the nucleotide exchange and eventually slow down the process of translation, causing vanishing white matter disease. We constructed a Saccharomyces cerevisiae genomic DNA library in YEp24 vector and screened it for the identification of extragenic suppressors of eIF2B mutations, corresponding to human eIF2B mutations. We found a suppressor-II (Sup-II) genomic clone, as suppressor of eIF2Bβ (gcd7-201) mutation. Identification of Sup-II reveals the presence of truncated SEC15, full-length TAN1 (tRNA acetyltransferase), full-length EMC4, full-length YGL230C (putative protein) and truncated SAP4 genes. Full-length TAN1 (tRNA acetyltransferase) gene, subcloned into pEG(KG) vector and overexpressed in gcd7-201 gcn2∆ strain, suppresses the slow-growth (Slg−) and general control derepression (Gcd−) phenotype of gcd7-201 gcn2∆ mutation, but YGL230C did not show any effect. A GST-Tan1p fusion protein of 60 kDa was detected by western blotting using α-GST antibodies. Interestingly, Tan1p overexpression also suppresses the temperature-sensitive (Ts−), Slg− and Gcd− phenotype of eIF2Bγ (gcd1-502) mutant. Role of Tan1p protein in eIF2B-mediated translation regulation was also studied. Results revealed that Tan1p overexpression confers resistance to GCD7 GCN2, gcd7-201 gcn2∆, GCD7 gcn2∆ growth defect under ethanol, H2O2 and caffeine stress. No resistance to DMSO-, NaCl- and DTT-mediated growth defect upon GCD7 gcn2∆, GCD7 GCN2, gcd7-201 gcn2∆ was observed by overexpression of TAN1. Hence, we proposed that Tan1p is involved directly or indirectly in regulating eIF2B-mediated translation. |
| Starting Page | 6464 |
| Ending Page | 6474 |
| Page Count | 11 |
| File Format | PDF HTM / HTML |
| PubMed reference number | 28677085v1 |
| Alternate Webpage(s) | https://doi.org/10.1007/s13205-017-0857-8 |
| DOI | 10.1007/s13205-017-0857-8 |
| Journal | 3 Biotech |
| Volume Number | 7 |
| Language | English |
| Access Restriction | Open |
| Subject Keyword | Acyltransferase Caffeine Clone Dithiothreitol EIF2AK4 wt Allele Ethanol Eukaryotic Initiation Factor-2 Eukaryotic Initiation Factor-2B F Factor GCD7 protein, S cerevisiae Genetic Translation Process Guanine Nucleotides Hydrogen Peroxide Leukoencephalopathies Mutation NOTCH1 protein, human Paget's Disease, Mammary Peptide Initiation Factors THUMPD1 gene Transcription Initiation Western Blotting White matter biological adaptation to stress |
| Content Type | Text |
| Resource Type | Article |
